CHO-Anti-Human PTP4A1 scFv stable cell line is clonally-derived from a CHO cell line, which has been transfected with an anti-human PTP4A1 scFv gene to allow expression of the scFv. It is an example of a cell line transfected using our proprietary CBTGS gene screening and amplification system.

Background:

Protein tyrosine phosphatase which stimulates progression from G1 into S phase during mitosis. May play a role in the development and maintenance of differentiating epithelial tissues. Enhances cell proliferation, cell motility and invasive activity, and promotes cancer metastasis.

1. Remove the packaging cell lines from liquid nitrogen and carry out a quick thaw. Float the cells in the 37 °C water bath for 2 minutes until nearly (80%) thawed. Once cells are thawed, it is important to dilute the cells 1:10 in growth media immediately to reduce the potentially toxic effects of the DMSO preservative on the cells.2. Clean the outside of the vial with 70% ethanol before opening.3. Remove the cells from the vial and add slowly into a 15 ml conical tube containing 10 ml pre-warmed media. 4. Centrifuge for 5 minutes 300 xg to pellet cells and remove the supernatant.5. Add 14 ml of media and transfer cells to a T25 shaker flask.6. Place the cells in the 37 °C incubator with 5% CO2.7. Allow incubation for 3-4 days to reach confluence. The cells will grow well over a period of several days in culture at 37 °C.

Subculturing:

1. Centrifuge for 5 minutes 300 xg and discard culture medium.2. Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.3. Add appropriate aliquots of the cell suspension to new culture vessels.Incubate cultures at 37 °C.Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:8 is recommended Medium Renewal: 2 to 3 times per week

Mycoplasma:

Mycoplasma Status: Negative (MycoAlert Kit)

Freeze Medium:

Complete growth medium 90%; DMSO, 10%

Storage:

Liquid nitrogen vapor phase

Preservation:

1. Detach cells from culture dish according to the Sub-Culture Procedure.2. Resuspend cells at a density of 5 x 10^6 cells/mL in freeze medium.3. Aliquot 1 mL cells into cryogenic vials.4. Place vials in a freezing container and store at –80 °C overnight.5. Transfer vials to liquid nitrogen for long term storage. If properly stored, cells should remain stable for years.

Safety Considerations:

The following safety precautions should be observed.1. Use pipette aids to prevent ingestion and keep aerosols down to a minimum.2. No eating, drinking or smoking while handling the stable line.3. Wash hands after handling the stable line and before leaving the lab.4. Decontaminate work surface with disinfectant or 70% ethanol before and after working with stable cells.5. All waste should be considered hazardous.6. Dispose of all liquid waste after each experiment and treat with bleach.

Ship:

Dry ice

Product Type:

scFv

Product Purity:

HPLC >98%, One strip for SDS-PAGE.

Product Activity/Specificity:

Tested positive against native human antigen.

Product Storage:

It should be stored at –20 °C. Reconstituted protein aliquots should avoid repeated freeze-thaw cycles.